In the field of cell culture, researchers are seeking cell culture surfaces which improve cell characteristics such as cell growth in culture. Reducing potentially contaminating ingredients such as serum or cell extracts is also desirable. Polymeric surfaces have been used for cell culture. In some cases, these polymeric surfaces have been formed from monomers containing amino acids or peptides. Acrylated or methacrylated amino acids have been used to form cell culture surfaces.
For example, Hern and Hubbell (Diane L. Hern and Jeffrey A. Hubbell; Incorporation of Adhesion Peptides into Nonadhesive Hydrogels useful for Tissue Resurfacing; J. Biomed. Mater. Res. 39, 266 (1998)) disclosed the formation of a (meth)acrylated peptide, including an adhesion peptide having an RGD sequence, by functionalizing the amine terminus of the peptide with an acrylate moiety. These functionalized peptides were then copolymerized with polyethylene glycol (PEG) or PEG diacrylate to form hydrogel cell culture surfaces having incorporated cell adhesion sequences.
Successful culture of difficult-to-culture cells requires that cell culture surfaces be tailored to accommodate the particular requirements of these cells. Bone cells, for example, prefer to be cultured in the presence of hydroxyapatite surfaces such as those discussed in Song et al. Song et al (Jie Song, Vienghkam Malathong, Carolyn R. Bertozzi, Mineralization of Synthetic Polymer Scaffolds: A Bottom-Up Approach for the Development of Artificial Bone, J. Am. Chem. Socl., 2005, 127, 3366-3372) disclosed the use of anionic groups such as methacrylated GLY (GlyMA), SER (SerMA), ASP (SerMA) and GLU (GluMA), and a methacrylated amino acid sequence RGD, a known cell adhesive monomer, to form a polymeric hydrogel containing 2-hydroxyethyl methacrylate (HEMA) or 2-hydroxyethyl dimethacrylamide (HEMAm) and cross-linkers ethylene glycol dimethacrylte (EGDMA) or ethylene glycol dimethacrylamide (EGDMAm). These anionic functionalized hydrogels were then used to provide a substrate for hydroxyapatite mineralization, after exposure of the hydrogel to urea. The negatively charged monomers, along with hydroxyethyl ester side chains of pHEMA that may have been hydrolyzed during the mineralization process, provided Ca2+ binding sites and allowed for the formation of a mineralized hydroxyapatite cell culture surface.
Ciucurel and Sefton (Ema C. Ciucurel and Michael V. Sefton; A Poloxamine-Polylysine Acrylate Scoffold for Modular Tissue Engineering; J. Biomaterials Science, 2010 DOI:10.1163/092050610X541133) disclosed the use of acrylated polylysine polymerized with poloxamine to form a poloxamine-polylysine acrylate (PPA) photopolymerized polymer. PPA hydrogels were able to support the proliferation of human microvascular endothelial cells (HMEC-1, a cell line) in culture.
Melkoumian et. al. (Jennifer L. Weber, David M. Weber, Andrei G. Fadeev, Yue Zhou, Paula Dolley-Sonneville, Jiwei Yang, Liqun Qui, Catherine A. Priest, Christopher Shogbon, Arthur W. Martin, Jodelle Nelson, Peter West, James P. Beltzer, Santona Pal and Ralph Brandenberger), Synthetic peptide-Acrylate Surfaces for Long-Term Self-Renewal and Cardiomyocyte Differentiation of Human Embryonic Stem Cells, (2010) Nature Biotechnology, Vol. 28, Number 6, 606-610 disclosed peptide-acrylate surface for long-term culture and differentiation of cardiomyocytes derived from human embryonic stem cells.